A. Materials and Methods · Web viewA. Materials and Methods2 B. Experimental Procedures3 C....

Supporting Information Page S1

UnyLinker Dimer Impurity Characterization and Process Improvement

Joshua L. Brooks, Phil Olsen, Lijian Chen, Andrew A. Rodriguez

Ionis Pharmaceuticals2855 Gazelle Court, Carlsbad, CA 92010

Supporting Information

A. Materials and Methods.....................................................................................................................................................2

B. Experimental Procedures..................................................................................................................................................3

C. Representative HPLC traces..............................................................................................................................................5

D. 1H and 13C NMR Spectra....................................................................................................................................................6

E. X-Ray Crystal Data...........................................................................................................................................................11


A. MATERIALS AND METHODS ReagentsReagents were obtained from Aldrich Chemical (www.sigma-aldrich.com). HPLC grade solvents were obtained from EMD Millipore (www.emdmillipore.com) and used as received.

ChromatographyTLC was performed on 0.25 mm E. Merck silica gel 60 F254 plates and visualized under UV light (254 nm) or by staining with potassium permanganate (KMnO4), cerium ammonium molybdenate (CAM), or p-anisaldehyde.

Analytical InstrumentationNMR spectra were recorded on a Bruker UltraShield Plus 300 MHz NMR at 24 °C in D6-DMSO unless otherwise indicated. Chemical shifts are expressed in ppm relative to solvent signals: D6-DMSO (13C, 39.52 ppm); coupling constants are expressed in Hz. NMR spectra were processed using ACD/SpecManger (www.acdlabs.com). LCMS spectra were on an Agilent Infinity Series LC and quadrupole MS by electrospray (ESI) ionization.

Compounds not cited in the paper are numbered herein from S1.

http://www.emdmillipore.com/


B. EXPERIMENTAL PROCEDURES

5,6-dihydroxy-2-phenylhexahydro-1H-4,7-epoxyisoindole-1,3(2H)-dione (9) and 6,6'-oxybis(5-hydroxy-2-phenylhexahydro-1H-4,7-epoxyisoindole-1,3(2H)-dione) (10): The Alkene 8 (10.00 g, 41.45 mmol) was suspended in 110 mL of acetone, and a 30% H2O2 (21.85 mL, 192.8 mmol, 4.65 equiv.). A solution of OsO4 in tBuOH (0.02 M, 10.36 mL, 0.01 equiv.) was then added and the reaction mixture brought to reflux. As the solution begins to reflux the alkene fully dissolves (~5 min.), the diol then begins to precipitate from solution (~15 min.). Reflux is continued for 4h at which point the solution is cooled to room temperature and quenched with sat. aqueous sodium thiosulfate (50 mL) and the diol collected by filtration. The solid was washed with H2O (50 mL) and acetone (100 mL), collected and dried under vacuum to yield a mixture of diol 9 and dimer 10 (7.84 g, containing 16% dimer).

Purification of Dimer 10 was accomplished by reaction of a mixture of diol 9 and 10 with dimethoxy propane and catalytic pTSA to give acetonide S1. The dimer was then purified by trituration.

6,6'-oxybis(5-hydroxy-2-phenylhexahydro-1H-4,7-epoxyisoindole-1,3(2H)-dione) (10): A crude mixture of diol 9 (10.00 g, 36.35 mmol) containing ~17% dimer 10 was suspended in 100 mL of dimethoxy propane and p-Toluenesulfonic acid monohydrate (69 mg, 0.364 mmol, 0.1 equiv.). The solution was then heated to 70 °C until the diol had been completely consumed as judged by HPLC. The solution was then cooled to room temperature and the remaining solid containing the dimer 10 was collected by filtration. The crude mixture was then purified by trituration with MeCN, to give the dimer 10 (1.68 g) as a white solid.

1H NMR (300 MHz) δ 7.55-7.38 (m, 6H), 7.20 (d, J = 6.0 Hz, 4H), 4.92 (broad s, 2H), 4.78 (s, 2H), 4.40 (s, 2H), 4.07 (d, J = 7.0 Hz, 2H), 3.94 (d, J = 7.0 Hz, 2H), 3.15 (q, J = 74.5 Hz, 4H). 13C NMR (75 MHz) δ 176.0, 132.2, 128.9, 126.7, 84.8, 82.0, 80.6, 73.0, 45.8.


2,2-dimethyl-6-phenylhexahydro-5H-4,8-epoxy[1,3]dioxolo[4,5-f]isoindole-5,7(6H)-dione (S1): 1H NMR (300 MHz) δ 8.02-7.38 (m, 2H), 7.24-7.18 (m, 2H), 4.57 (s, 2H) 4.53 (s, 2H), 3.14 (s, 2H), 1.35 (s, 3H), 1.23 (s, 3H). 13C NMR (75 MHz) δ 175.8, 132.2, 128.9, 126.8, 111.1, 81.0, 80.6, 44.8, 25.7, 25.0.

Oxybis(1,3-dioxo-2-phenyloctahydro-1H-4,7-epoxyisoindole-6,5-diyl) bis(4-nitrobenzoate): In a 10 mL roundbottom flask containing dimer diol 10 (100 mg, 0.188 mmol) suspended in pyridine (5 mL) was added 4-Nitrobenzoyl chloride (348 mg, 1.90 mmol), and the suspension stirred at room temperature under a nitrogen atmosphere until complete conversion had occurred as judged by TLC. The reaction mixture was then quenched with 10% HCl (10 mL) and extracted with CH 2Cl2 (3 x 25 mL), washed with 5% NaOH, dried (MgSO4), filtered, and concentrated by rotary evaporation to afford the crude product as a white powder. The product was recrystallized from MeCN to give the bis- nitrobenzoate (122 mg, 78% yield).

1H NMR (300 MHz) δ 8.17 (d, J = 8.4 Hz, 2H), 7.87 (d, J = 8.8 Hz, 2H), 7.55-7.41 (m, 3H), 7.27-7.22 (m 2H), 5.27 (d, J = 6.7 Hz, 1H), 4.82 (s, 1H), 4.65 (s, 1H), 4.44 (d, J = 5.9 Hz, 1H), 3.39 (d, J = 5.6 Hz, 1H), 3.30 (d, J = 5.6 Hz, 1H). 13C NMR (75 MHz) δ 166.9, 165.76, 164.8, 150.2, 150.0, 136.4, 135.0, 131.7, 131.5, 130.7, 130.6, 128.6, 123.8, 123.7, 67.4, 52.8, 38.1, 29.8, 28.4, 23.2, 22.4, 13.8, 10.8.

4-phenylhexahydro-3H-2,6-epoxyoxireno[2,3-f]isoindole-3,5(4H)-dione (11): The Alkene 8 (1.00 g, 4.14 mmol) was dissolved in 50 mL of CH2Cl2, and 3-Chloroperbenzoic acid (1.43 g, 8.29 mmol, 2 equiv.), the solution was stirred at room temperature until complete conversion had occurred as judged by TLC. The reaction mixture was then quenched with sat. aqueous sodium thiosulfate (25 mL) and extracted with CH2Cl2 (3 x 25 mL), washed with 5% NaOH, dried (MgSO4), filtered, and concentrated by rotary evaporation to afford the epoxide 11 as a white powder without need for further purification.

1H NMR (300 MHz) δ 7.53 – 7.40 (m, 3H), 7.22 – 7.18 (m, 2H), 4.77 (s, 2H), 3.75 (s, 2H), 3.33 (d, J = 4.35, 2H). 13C NMR (75 MHz) δ 175.5, 132.1, 129.0, 128.5, 126.8, 76.5, 49.3, 47.9.


C. REPRESENTATIVE HPLC TRACES

Figure S1: Representative HPLC traces for dihydroxylation of alkene 8 to diol 9 and dimer 10. A: H2O2 used as co-oxidant. B: NMO used as the co-oxidant.

A

B


D. 1 H AND 13 C NMR SPECTRA


E. X-RAY CRYSTAL DATA

Table 1. Crystal data and structure refinement for ISIS22_a.Identification code isis22_aEmpirical formula C44 H33 N5 O15Formula weight 871.75Temperature 100.0 KWavelength 1.54178 ÅCrystal system MonoclinicSpace group P 1 21/c 1Unit cell dimensions a = 11.0713(3) Å = 90°.

b = 12.1881(3) Å = 96.882(2)°.c = 30.0378(7) Å = 90°.

Volume 4024.04(18) Å3

Z 4Density (calculated) 1.439 Mg/m3

Absorption coefficient 0.935 mm-1

F(000) 1808Crystal size 0.22 x 0.1 x 0.07 mm3

Theta range for data collection 2.964 to 68.253°.Index ranges -13<=h<=10, -13<=k<=14, -35<=l<=35Reflections collected 32626Independent reflections 7281 [R(int) = 0.0272]Completeness to theta = 67.500° 99.5 % Absorption correction Semi-empirical from equivalentsMax. and min. transmission 0.1665 and 0.0681Refinement method Full-matrix least-squares on F2

Data / restraints / parameters 7281 / 0 / 578Goodness-of-fit on F2 1.022Final R indices [I>2sigma(I)] R1 = 0.0346, wR2 = 0.0891R indices (all data) R1 = 0.0379, wR2 = 0.0916Extinction coefficient n/aLargest diff. peak and hole 0.207 and -0.197 e.Å-3


Table 2. Atomic coordinates ( x 104) and equivalent isotropic displacement parameters (Å2x 103)for ISIS22_a. U(eq) is defined as one third of the trace of the orthogonalized Uij tensor.________________________________________________________________________________

x y z U(eq)________________________________________________________________________________ O(1) -1069(1) 2418(1) 3549(1) 38(1)O(2) 2379(1) 461(1) 4004(1) 42(1)O(3) 1965(1) 3200(1) 3310(1) 35(1)O(4) 2177(1) 5600(1) 3466(1) 34(1)O(5) 3187(1) 6618(1) 4016(1) 41(1)O(6) 2392(1) 10987(1) 2536(1) 55(1)O(7) 1220(1) 9957(1) 2087(1) 55(1)O(8) 3140(2) 4426(1) 703(1) 72(1)O(9) 3111(1) 6101(1) 918(1) 63(1)O(10) 4175(1) 4486(1) 3601(1) 34(1)O(11) 5027(1) 2881(1) 2904(1) 40(1)O(12) 5648(1) 4635(1) 3001(1) 33(1)O(13) 6281(1) 5964(1) 3833(1) 33(1)O(14) 7235(1) 5767(1) 5081(1) 47(1)O(15) 9783(1) 5828(1) 3998(1) 43(1)N(1) 543(1) 1222(1) 3710(1) 36(1)N(2) 1937(1) 10100(1) 2426(1) 44(1)N(3) 3272(1) 5127(1) 994(1) 49(1)N(4) 8627(1) 6028(1) 4581(1) 35(1)C(1) -16(2) -717(1) 3731(1) 48(1)C(2) -491(2) -1651(1) 3505(1) 58(1)C(3) -966(2) -1594(2) 3060(1) 60(1)C(4) -976(2) -608(2) 2832(1) 54(1)C(5) -485(1) 323(1) 3049(1) 44(1)C(6) -10(1) 260(1) 3497(1) 39(1)C(7) -28(1) 2244(1) 3706(1) 34(1)C(8) 879(1) 3076(1) 3918(1) 34(1)C(9) 2068(1) 2432(1) 4028(1) 34(1)C(10) 1739(1) 1254(1) 3925(1) 36(1)C(11) 1213(1) 3882(1) 3557(1) 34(1)C(12) 2105(1) 4726(1) 3781(1) 33(1)C(13) 3301(1) 4050(1) 3861(1) 33(1)C(14) 2852(1) 2923(1) 3683(1) 34(1)


C(15) 2729(1) 6516(1) 3634(1) 35(1)C(16) 2642(1) 7410(1) 3291(1) 35(1)C(17) 3002(1) 8455(1) 3432(1) 40(1)C(18) 2813(1) 9335(1) 3142(1) 41(1)C(19) 2255(1) 9146(1) 2717(1) 38(1)C(20) 1944(2) 8106(1) 2557(1) 48(1)C(21) 2151(2) 7233(1) 2848(1) 46(1)C(22) 5369(1) 4236(1) 3795(1) 32(1)C(23) 5866(1) 5142(1) 4124(1) 33(1)C(24) 7070(1) 4734(1) 4382(1) 35(1)C(25) 7925(1) 4774(1) 4014(1) 35(1)C(26) 7074(1) 5265(1) 3616(1) 34(1)C(27) 6256(1) 4326(1) 3431(1) 33(1)C(28) 5056(1) 3809(1) 2768(1) 34(1)C(29) 4514(1) 4176(1) 2315(1) 35(1)C(30) 4385(1) 5283(1) 2207(1) 40(1)C(31) 3945(1) 5594(1) 1774(1) 42(1)C(32) 3672(1) 4782(1) 1459(1) 41(1)C(33) 3780(1) 3675(1) 1556(1) 43(1)C(34) 4189(1) 3377(1) 1992(1) 39(1)C(35) 8907(1) 5580(1) 4178(1) 36(1)C(36) 7599(1) 5552(1) 4730(1) 37(1)C(37) 9296(1) 6908(1) 4809(1) 36(1)C(38) 10539(1) 6816(1) 4929(1) 40(1)C(39) 11159(1) 7678(1) 5155(1) 43(1)C(40) 10547(2) 8616(1) 5257(1) 43(1)C(41) 9306(1) 8707(1) 5125(1) 41(1)C(42) 8677(1) 7853(1) 4901(1) 38(1)N(5) 5049(2) -2388(2) 5335(1) 105(1)C(43) 3931(2) -1134(2) 4730(1) 66(1)C(44) 4577(2) -1837(2) 5067(1) 62(1)________________________________________________________________________________


Table 3. Bond lengths [Å] and angles [°] for ISIS22_a._____________________________________________________ O(1)-C(7) 1.2098(17)O(2)-C(10) 1.2053(17)O(3)-C(11) 1.4431(16)O(3)-C(14) 1.4407(16)O(4)-C(12) 1.4319(16)O(4)-C(15) 1.3420(17)O(5)-C(15) 1.2051(17)O(6)-N(2) 1.2214(18)O(7)-N(2) 1.2267(17)O(8)-N(3) 1.2181(19)O(9)-N(3) 1.2179(19)O(10)-C(13) 1.4181(16)O(10)-C(22) 1.4126(16)O(11)-C(28) 1.2047(17)O(12)-C(27) 1.4346(16)O(12)-C(28) 1.3494(16)O(13)-C(23) 1.4410(16)O(13)-C(26) 1.4350(16)O(14)-C(36) 1.2036(18)O(15)-C(35) 1.2041(18)N(1)-C(6) 1.4364(18)N(1)-C(7) 1.3969(18)N(1)-C(10) 1.4024(18)N(2)-C(19) 1.4719(19)N(3)-C(32) 1.4756(19)N(4)-C(35) 1.3961(18)N(4)-C(36) 1.3976(19)N(4)-C(37) 1.4309(18)C(1)-H(1) 0.9500C(1)-C(2) 1.396(2)C(1)-C(6) 1.383(2)C(2)-H(2) 0.9500C(2)-C(3) 1.377(3)C(3)-H(3) 0.9500C(3)-C(4) 1.382(3)C(4)-H(4) 0.9500


C(4)-C(5) 1.386(2)C(5)-H(5) 0.9500C(5)-C(6) 1.387(2)C(7)-C(8) 1.5132(19)C(8)-H(8) 1.0000C(8)-C(9) 1.5340(19)C(8)-C(11) 1.5408(19)C(9)-H(9) 1.0000C(9)-C(10) 1.5045(19)C(9)-C(14) 1.5486(18)C(11)-H(11) 1.0000C(11)-C(12) 1.5252(19)C(12)-H(12) 1.0000C(12)-C(13) 1.5540(18)C(13)-H(13) 1.0000C(13)-C(14) 1.5352(19)C(14)-H(14) 1.0000C(15)-C(16) 1.4930(19)C(16)-C(17) 1.386(2)C(16)-C(21) 1.392(2)C(17)-H(17) 0.9500C(17)-C(18) 1.381(2)C(18)-H(18) 0.9500C(18)-C(19) 1.369(2)C(19)-C(20) 1.384(2)C(20)-H(20) 0.9500C(20)-C(21) 1.378(2)C(21)-H(21) 0.9500C(22)-H(22) 1.0000C(22)-C(23) 1.5389(19)C(22)-C(27) 1.5577(18)C(23)-H(23) 1.0000C(23)-C(24) 1.5405(18)C(24)-H(24) 1.0000C(24)-C(25) 1.5396(19)C(24)-C(36) 1.511(2)C(25)-H(25) 1.0000C(25)-C(26) 1.5497(19)


C(25)-C(35) 1.504(2)C(26)-H(26) 1.0000C(26)-C(27) 1.5224(19)C(27)-H(27) 1.0000C(28)-C(29) 1.4904(19)C(29)-C(30) 1.390(2)C(29)-C(34) 1.391(2)C(30)-H(30) 0.9500C(30)-C(31) 1.385(2)C(31)-H(31) 0.9500C(31)-C(32) 1.378(2)C(32)-C(33) 1.384(2)C(33)-H(33) 0.9500C(33)-C(34) 1.381(2)C(34)-H(34) 0.9500C(37)-C(38) 1.385(2)C(37)-C(42) 1.385(2)C(38)-H(38) 0.9500C(38)-C(39) 1.387(2)C(39)-H(39) 0.9500C(39)-C(40) 1.381(2)C(40)-H(40) 0.9500C(40)-C(41) 1.388(2)C(41)-H(41) 0.9500C(41)-C(42) 1.382(2)C(42)-H(42) 0.9500N(5)-C(44) 1.128(3)C(43)-H(43A) 0.9800C(43)-H(43B) 0.9800C(43)-H(43C) 0.9800C(43)-C(44) 1.447(3)

C(14)-O(3)-C(11) 96.60(9)C(15)-O(4)-C(12) 115.54(10)C(22)-O(10)-C(13) 111.01(10)C(28)-O(12)-C(27) 114.67(10)C(26)-O(13)-C(23) 96.87(9)C(7)-N(1)-C(6) 123.72(12)


C(7)-N(1)-C(10) 112.36(11)C(10)-N(1)-C(6) 123.84(12)O(6)-N(2)-O(7) 123.65(13)O(6)-N(2)-C(19) 118.52(13)O(7)-N(2)-C(19) 117.83(13)O(8)-N(3)-C(32) 118.49(14)O(9)-N(3)-O(8) 123.14(14)O(9)-N(3)-C(32) 118.36(13)C(35)-N(4)-C(36) 112.90(12)C(35)-N(4)-C(37) 123.54(12)C(36)-N(4)-C(37) 123.53(11)C(2)-C(1)-H(1) 120.7C(6)-C(1)-H(1) 120.7C(6)-C(1)-C(2) 118.61(17)C(1)-C(2)-H(2) 119.7C(3)-C(2)-C(1) 120.63(17)C(3)-C(2)-H(2) 119.7C(2)-C(3)-H(3) 119.9C(2)-C(3)-C(4) 120.20(16)C(4)-C(3)-H(3) 119.9C(3)-C(4)-H(4) 120.0C(3)-C(4)-C(5) 120.00(17)C(5)-C(4)-H(4) 120.0C(4)-C(5)-H(5) 120.3C(4)-C(5)-C(6) 119.46(16)C(6)-C(5)-H(5) 120.3C(1)-C(6)-N(1) 120.27(14)C(1)-C(6)-C(5) 121.08(14)C(5)-C(6)-N(1) 118.58(13)O(1)-C(7)-N(1) 124.71(13)O(1)-C(7)-C(8) 126.81(13)N(1)-C(7)-C(8) 108.47(11)C(7)-C(8)-H(8) 113.2C(7)-C(8)-C(9) 104.92(11)C(7)-C(8)-C(11) 109.45(11)C(9)-C(8)-H(8) 113.2C(9)-C(8)-C(11) 101.88(10)C(11)-C(8)-H(8) 113.2


C(8)-C(9)-H(9) 112.6C(8)-C(9)-C(14) 101.13(10)C(10)-C(9)-C(8) 105.26(11)C(10)-C(9)-H(9) 112.6C(10)-C(9)-C(14) 111.83(11)C(14)-C(9)-H(9) 112.6O(2)-C(10)-N(1) 124.65(13)O(2)-C(10)-C(9) 127.05(13)N(1)-C(10)-C(9) 108.29(11)O(3)-C(11)-C(8) 101.39(10)O(3)-C(11)-H(11) 114.1O(3)-C(11)-C(12) 103.42(10)C(8)-C(11)-H(11) 114.1C(12)-C(11)-C(8) 108.50(11)C(12)-C(11)-H(11) 114.1O(4)-C(12)-C(11) 107.20(10)O(4)-C(12)-H(12) 111.6O(4)-C(12)-C(13) 112.54(10)C(11)-C(12)-H(12) 111.6C(11)-C(12)-C(13) 101.89(10)C(13)-C(12)-H(12) 111.6O(10)-C(13)-C(12) 109.91(10)O(10)-C(13)-H(13) 111.6O(10)-C(13)-C(14) 111.00(11)C(12)-C(13)-H(13) 111.6C(14)-C(13)-C(12) 100.66(10)C(14)-C(13)-H(13) 111.6O(3)-C(14)-C(9) 103.18(10)O(3)-C(14)-C(13) 102.92(10)O(3)-C(14)-H(14) 114.1C(9)-C(14)-H(14) 114.1C(13)-C(14)-C(9) 107.30(11)C(13)-C(14)-H(14) 114.1O(4)-C(15)-C(16) 111.10(11)O(5)-C(15)-O(4) 124.33(13)O(5)-C(15)-C(16) 124.49(13)C(17)-C(16)-C(15) 117.96(12)C(17)-C(16)-C(21) 119.83(13)


C(21)-C(16)-C(15) 122.10(13)C(16)-C(17)-H(17) 119.8C(18)-C(17)-C(16) 120.38(13)C(18)-C(17)-H(17) 119.8C(17)-C(18)-H(18) 120.8C(19)-C(18)-C(17) 118.32(14)C(19)-C(18)-H(18) 120.8C(18)-C(19)-N(2) 118.03(13)C(18)-C(19)-C(20) 122.88(14)C(20)-C(19)-N(2) 119.06(13)C(19)-C(20)-H(20) 120.9C(21)-C(20)-C(19) 118.12(14)C(21)-C(20)-H(20) 120.9C(16)-C(21)-H(21) 119.9C(20)-C(21)-C(16) 120.23(14)C(20)-C(21)-H(21) 119.9O(10)-C(22)-H(22) 111.7O(10)-C(22)-C(23) 110.79(11)O(10)-C(22)-C(27) 109.45(10)C(23)-C(22)-H(22) 111.7C(23)-C(22)-C(27) 101.16(10)C(27)-C(22)-H(22) 111.7O(13)-C(23)-C(22) 102.96(10)O(13)-C(23)-H(23) 114.1O(13)-C(23)-C(24) 102.07(10)C(22)-C(23)-H(23) 114.1C(22)-C(23)-C(24) 108.18(11)C(24)-C(23)-H(23) 114.1C(23)-C(24)-H(24) 112.6C(25)-C(24)-C(23) 101.50(10)C(25)-C(24)-H(24) 112.6C(36)-C(24)-C(23) 111.98(11)C(36)-C(24)-H(24) 112.6C(36)-C(24)-C(25) 104.78(11)C(24)-C(25)-H(25) 112.9C(24)-C(25)-C(26) 101.29(10)C(26)-C(25)-H(25) 112.9C(35)-C(25)-C(24) 105.53(11)


C(35)-C(25)-H(25) 112.9C(35)-C(25)-C(26) 110.43(11)O(13)-C(26)-C(25) 103.17(10)O(13)-C(26)-H(26) 114.2O(13)-C(26)-C(27) 104.03(10)C(25)-C(26)-H(26) 114.2C(27)-C(26)-C(25) 105.71(11)C(27)-C(26)-H(26) 114.2O(12)-C(27)-C(22) 112.56(10)O(12)-C(27)-C(26) 109.01(11)O(12)-C(27)-H(27) 111.2C(22)-C(27)-H(27) 111.2C(26)-C(27)-C(22) 101.22(10)C(26)-C(27)-H(27) 111.2O(11)-C(28)-O(12) 123.78(12)O(11)-C(28)-C(29) 124.60(13)O(12)-C(28)-C(29) 111.52(11)C(30)-C(29)-C(28) 121.53(12)C(30)-C(29)-C(34) 120.43(13)C(34)-C(29)-C(28) 117.97(13)C(29)-C(30)-H(30) 120.0C(31)-C(30)-C(29) 119.93(13)C(31)-C(30)-H(30) 120.0C(30)-C(31)-H(31) 120.9C(32)-C(31)-C(30) 118.14(14)C(32)-C(31)-H(31) 120.9C(31)-C(32)-N(3) 117.58(14)C(31)-C(32)-C(33) 123.34(14)C(33)-C(32)-N(3) 119.06(13)C(32)-C(33)-H(33) 121.1C(34)-C(33)-C(32) 117.79(14)C(34)-C(33)-H(33) 121.1C(29)-C(34)-H(34) 119.8C(33)-C(34)-C(29) 120.30(14)C(33)-C(34)-H(34) 119.8O(15)-C(35)-N(4) 124.43(13)O(15)-C(35)-C(25) 127.32(13)N(4)-C(35)-C(25) 108.24(12)


O(14)-C(36)-N(4) 124.17(13)O(14)-C(36)-C(24) 127.46(13)N(4)-C(36)-C(24) 108.37(11)C(38)-C(37)-N(4) 120.36(13)C(42)-C(37)-N(4) 118.62(12)C(42)-C(37)-C(38) 121.02(13)C(37)-C(38)-H(38) 120.5C(37)-C(38)-C(39) 118.95(14)C(39)-C(38)-H(38) 120.5C(38)-C(39)-H(39) 119.7C(40)-C(39)-C(38) 120.55(14)C(40)-C(39)-H(39) 119.7C(39)-C(40)-H(40) 120.1C(39)-C(40)-C(41) 119.88(14)C(41)-C(40)-H(40) 120.1C(40)-C(41)-H(41) 119.9C(42)-C(41)-C(40) 120.18(14)C(42)-C(41)-H(41) 119.9C(37)-C(42)-H(42) 120.3C(41)-C(42)-C(37) 119.40(14)C(41)-C(42)-H(42) 120.3H(43A)-C(43)-H(43B) 109.5H(43A)-C(43)-H(43C) 109.5H(43B)-C(43)-H(43C) 109.5C(44)-C(43)-H(43A) 109.5C(44)-C(43)-H(43B) 109.5C(44)-C(43)-H(43C) 109.5N(5)-C(44)-C(43) 177.9(2)_____________________________________________________________ Symmetry transformations used to generate equivalent atoms:


Table 4. Anisotropic displacement parameters (Å2x 103) for ISIS22_a. The anisotropicdisplacement factor exponent takes the form: -22[ h2 a*2U11 + ... + 2 h k a* b* U12 ]______________________________________________________________________________

U11 U22 U33 U23 U13 U12

______________________________________________________________________________ O(1) 32(1) 39(1) 44(1) -1(1) 4(1) 1(1)O(2) 42(1) 36(1) 46(1) 2(1) 4(1) 6(1)O(3) 34(1) 36(1) 34(1) -1(1) 4(1) 1(1)O(4) 34(1) 33(1) 36(1) 1(1) 3(1) 0(1)O(5) 45(1) 39(1) 36(1) 0(1) -1(1) -2(1)O(6) 70(1) 39(1) 56(1) 6(1) 7(1) 4(1)O(7) 44(1) 72(1) 48(1) 21(1) -2(1) -5(1)O(8) 106(1) 67(1) 38(1) -5(1) -14(1) -21(1)O(9) 78(1) 62(1) 46(1) 11(1) -7(1) 5(1)O(10) 29(1) 39(1) 33(1) 3(1) 3(1) 1(1)O(11) 45(1) 34(1) 41(1) 1(1) 0(1) 0(1)O(12) 34(1) 34(1) 30(1) -1(1) 2(1) -2(1)O(13) 34(1) 33(1) 33(1) 1(1) 4(1) 2(1)O(14) 51(1) 57(1) 32(1) -4(1) 8(1) -10(1)O(15) 32(1) 52(1) 44(1) -5(1) 6(1) 0(1)N(1) 36(1) 33(1) 39(1) 0(1) 6(1) 0(1)N(2) 40(1) 48(1) 44(1) 9(1) 10(1) 3(1)N(3) 48(1) 57(1) 39(1) 3(1) -4(1) -9(1)N(4) 33(1) 38(1) 34(1) 0(1) 1(1) 1(1)C(1) 44(1) 40(1) 60(1) 2(1) 11(1) -1(1)C(2) 51(1) 37(1) 89(1) -2(1) 18(1) -5(1)C(3) 48(1) 47(1) 87(1) -23(1) 17(1) -9(1)C(4) 45(1) 57(1) 60(1) -20(1) 10(1) -4(1)C(5) 40(1) 43(1) 49(1) -7(1) 9(1) -1(1)C(6) 34(1) 35(1) 49(1) -5(1) 10(1) -1(1)C(7) 36(1) 35(1) 34(1) 1(1) 8(1) 1(1)C(8) 33(1) 35(1) 35(1) -1(1) 5(1) 1(1)C(9) 34(1) 34(1) 34(1) 1(1) 4(1) 1(1)C(10) 37(1) 38(1) 34(1) 2(1) 7(1) 2(1)C(11) 31(1) 34(1) 35(1) 0(1) 4(1) 3(1)C(12) 32(1) 33(1) 33(1) 2(1) 4(1) 1(1)C(13) 31(1) 36(1) 31(1) 2(1) 5(1) 1(1)C(14) 32(1) 36(1) 34(1) 2(1) 4(1) 4(1)


C(15) 32(1) 36(1) 38(1) -2(1) 5(1) 2(1)C(16) 32(1) 37(1) 38(1) 0(1) 5(1) 2(1)C(17) 44(1) 40(1) 35(1) -1(1) 4(1) 0(1)C(18) 46(1) 36(1) 41(1) -1(1) 9(1) -1(1)C(19) 36(1) 41(1) 39(1) 6(1) 8(1) 2(1)C(20) 52(1) 51(1) 38(1) 4(1) -4(1) -9(1)C(21) 55(1) 40(1) 42(1) 0(1) -1(1) -8(1)C(22) 30(1) 34(1) 32(1) 2(1) 2(1) 2(1)C(23) 33(1) 36(1) 32(1) 2(1) 5(1) 1(1)C(24) 35(1) 36(1) 33(1) 3(1) 1(1) 0(1)C(25) 32(1) 35(1) 38(1) -2(1) 2(1) 3(1)C(26) 32(1) 37(1) 32(1) -1(1) 5(1) 1(1)C(27) 32(1) 35(1) 32(1) 1(1) 2(1) 2(1)C(28) 32(1) 34(1) 36(1) -2(1) 5(1) 1(1)C(29) 32(1) 38(1) 36(1) -2(1) 4(1) -1(1)C(30) 43(1) 37(1) 37(1) -4(1) -1(1) -1(1)C(31) 47(1) 38(1) 42(1) 2(1) -2(1) -1(1)C(32) 38(1) 49(1) 34(1) 2(1) -2(1) -5(1)C(33) 44(1) 45(1) 39(1) -7(1) 1(1) -7(1)C(34) 40(1) 36(1) 40(1) -2(1) 3(1) -3(1)C(35) 32(1) 38(1) 37(1) 1(1) -1(1) 6(1)C(36) 37(1) 39(1) 33(1) 5(1) -1(1) 1(1)C(37) 38(1) 37(1) 31(1) 2(1) 1(1) -2(1)C(38) 40(1) 39(1) 40(1) 2(1) 0(1) 2(1)C(39) 38(1) 44(1) 46(1) 5(1) -3(1) -4(1)C(40) 52(1) 37(1) 38(1) 3(1) 0(1) -7(1)C(41) 50(1) 35(1) 38(1) 3(1) 7(1) 1(1)C(42) 38(1) 40(1) 35(1) 5(1) 3(1) 2(1)N(5) 73(1) 139(2) 106(2) 67(2) 17(1) 34(1)C(43) 76(1) 68(1) 50(1) 9(1) -4(1) -5(1)C(44) 54(1) 75(1) 60(1) 16(1) 12(1) 8(1)______________________________________________________________________________


Table 5. Hydrogen coordinates ( x 104) and isotropic displacement parameters (Å2x 10 3)for ISIS22_a.________________________________________________________________________________

x y z U(eq)________________________________________________________________________________ H(1) 296 -752 4039 57H(2) -486 -2332 3659 70H(3) -1288 -2235 2910 72H(4) -1319 -569 2527 65H(5) -474 999 2892 52H(8) 606 3450 4185 41H(9) 2442 2540 4345 41H(11) 495 4208 3370 40H(12) 1846 4998 4069 40H(13) 3621 4018 4187 39H(14) 3513 2426 3605 41H(17) 3382 8568 3729 48H(18) 3064 10053 3235 49H(20) 1597 7996 2256 58H(21) 1958 6509 2747 55H(22) 5413 3497 3941 39H(23) 5265 5417 4321 40H(24) 7000 3981 4508 42H(25) 8253 4037 3945 42H(26) 7509 5644 3387 40H(27) 6731 3633 3412 40H(30) 4598 5825 2430 47H(31) 3835 6346 1697 51H(33) 3579 3136 1331 51H(34) 4249 2623 2072 47H(38) 10961 6172 4858 48H(39) 12011 7623 5241 52H(40) 10975 9197 5417 51H(41) 8888 9358 5189 49H(42) 7827 7914 4810 45H(43A) 3341 -1571 4536 98H(43B) 4512 -796 4550 98


H(43C) 3503 -559 4877 98________________________________________________________________________________

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